GSM7384900: BJ-HRAS-CRISPRi Clone b, D0; Homo sapiens; OTHER
Genomic DNA was isolated using the MasterPure Complete DNA Purification kit (Lucigen). Sequencing libraries were generated with two sequential rounds of PCR using Herculase II Fusion DNA polymerase (Agilent). 6.6 µg of genomic DNA was used in 100 µl reaction for the first round of PCR. A total of 40 reactions were performed per replicate and pooled following 18 cycles of PCR1 amplification. 5 µl from the pooled reaction product and primers containing barcodes and Illumina sequencing adaptors were used in the second round of PCR (11 – 12 cycles). CRISPR screen custom DNA library